The aim of this experimental study was to evaluate histomorphometrically the effects of light-emitting diode (LED) photobiomodulation therapy (LPT) on bone formation in response to expansion of the interpremaxillary suture in rats. Twenty male, 50- to 60-day-old Wistar rats were divided into two equal groups (control and experimental). Both groups were subjected to expansion for 5 days, and 50 cN of force was applied to the maxillary incisors with helical spring. An OsseoPulse® LED device, 618-nm wavelength and 20-mW/cm(2) output power irradiation, was applied to the interpremaxillary suture for 10 days. Bone formation in the sutural area was histomorphometrically evaluated, including the amount of new bone formation (in square micrometers), number of osteoblasts, number of osteoclasts, and number of vessels. Mann-Whitney U test was used for statistical evaluation at p < 0.025 level. Significant differences were found between groups for all investigated histomorphometric parameters. New bone formation area (p = 0.024, 1.48-fold), number of osteoblasts (p < 0.001, 1.59-fold), number of osteoclasts (p = 0.004, 1.43-fold), and number of vessels (p = 0.007, 1.67-fold) showed higher values in the experimental group than the control. Bone histomorphometric measurements revealed that bone architecture in the LPT group was improved. The application of LPT can stimulate bone formation in the orthopedically expanded interpremaxillary suture during expansion and the early phase of the retention periods.